Bulk segregant analysis was used to search for RAPD (random amplified polymorphic DNA) markers linked to gene(s) affecting oleic acid concentration in an F2 population from the Brassica rapa ssp. oleifera cross Jo4002 x a high oleic acid individual from line Jo4072. Eight primers (=8 markers) out of 104 discriminated the 'high' and 'low' bulks consisting of extreme individuals from the oleic acid distribution. These markers were analysed throughout the entire F2 population, and their association with oleic acid was studied using both interval mapping and ANOVA analysis. Six of the markers mapped to one linkage group. A quantitative trait locus (QTL) affecting oleic acid concentration was found to reside within this linkage group with a LOD score >15. The most suitable marker for oleic acid content is OPH-17, a codominant marker close (<4cM) to the QTL. The mean seed oleic acid content in the F2 individuals carrying the larger allele of this marker was 80.14±9.76%; in individuals with the smaller allele, 54.53±6.83%; in the heterozygotes, 65.47±8.15%. To increase reproducibility, the RAPD marker was converted into a SCAR (sequence characterized amplied region) marker with specific primers. Marker OPH-17 can be used to select spring turnip rape individuals with the desired oleic acid content.

The F2 progeny (64 individuals) from the cross between oilseed rape (Brassica napus L.) cultivar Topas and R4 (a low linolenic mutation line) was analyzed with 8 RFLPs and 34 RAPDs to discover a genetic tag for gene(s) affecting linolenic acid concentration. According to variance analysis (ANOVA), one RAPD marker (25a) was significantly associated with linolenic acid content; the linolenic acid concentration in the seeds of F2 individuals showing the marker (includes both homo- and hetero-zygotes) was 7.43 +/- 1.35% and in those lacking the marker was 5.70 +/- 1.52%. Marker 25a may be used to facilitate selection for fatty acid composition in future breeding programs of oilseed rape.

A linkage map of spring turnip rape (Brassica rapa ssp. oleifera) was conslructed from an F2 population of a cross Jo4002 x Sv3402. The map contained 22 RFLP loci, 144 RAPDs, one microsatellite, and one morphological marker (seed colour). All ten B.rapa linkage groups could be identified and the total map distance was 519 cM. A proportion of the markers (13%), most of which were located in two linkage groups, showed segregation distortion.

F2 progeny (105 individuals) from the cross Jo4002 x Sv3402 were used to identify DNA markers associated with palmitic-acid content in spring turnip rape (Brassica rapa ssp. oleifera). QTL mapping and ANOVA analysis of 140 markers exposed one linkage group with a locus controlling palmitic-acid content (LOD score 27), and one RAPD (random amplified polymorphic DNA) marker, OPB-11a, closely linked (1.4 cM) to this locus. Palmitic-acid content in the 62 F2 plants with the visible allele of marker OPB-11a was 8.45 ±3.15%, while that in the 24 plants without it was 4.59 ±0.97%. As oleic-acid concentration is affected by a locus on the same linkage group as the palmitic-acid locus, this locus probably controls the chain elongation from palmitic acid to oleic acid (through stearic acid). Marker OPB-11a may be used in future breeding programs of spring turnip rape to simplify and hasten the selection for palmitic-acid content.

Protoplast cultures were prepared from 6-day-old hypocotyls of six spring, seven winter cultivars of Brassica napus L. and one line of Brassica campestris L. The molarity of enzyme solution was raised to 0,714 M mannitol resulting in well manipulable, cytoplasm dense protoplasts. In the protoplast purification procedure density gradient centrifugation was used to minimize physical damage of protoplasts. Three different protoplast culture systems —(1) liquid, (2) 2nd day embedded, (3) directly embedded in low melting agarose were compared. The two different protoplast embedding techniques resulted in the same efficiency of cell division as the liquid culture method and over this fact the colony browning was avoided. Using protoplast agarose-embedding and culture techniques, healthy calli were obtained for plant regeneration experiments. Incorporation of silver nitrate into the regeneration medium improved the efficiency of plant regeneration in responsive genotypes and the regeneration was induced in three nonresponsive (without silver nitrate) genotypes, too. The supplement of silver nitrate in regeneration medium was especially advantageous in plant regeneration of B. campestris. Out of fourteen commercial cultivars of Brassica napus and B. campestris, there is only one recalcitrant genotype in obtaining plantlets from protoplast-derived calli.